Truffle Fraud Zoe Balestri Neeky Yassari Isabella Zappi

Truffle Fraud? Zoe Balestri, Neeky Yassari, Isabella Zappi, Joelle Windmiller Mentor: Howard Waldman Abstract: The purpose of this lab is to analyze the genetic sequence of a specific gene on Tuber Melanosporum, Tuber Sinensis, Tuber Indicum, and Tuber Himalayensis truffles to determine if there is fraud in the truffle industry. This was accomplished by examining the genetic makeup of different truffle samples through DNA sequencing. Unfortunately, accurate DNA results were not acquired, due to a unique trend specific to fungi. Materials & Methods: ● ● Criteria: Species and Price When: December 2015 Where: Internet and donated from teacher What: ‘Perigord’ French Black Truffle (Tuber Melanosporum) and Chinese Black Truffle (Tuber Indicum) ● How: Mail ● Number of Samples: 4 ● Primer used: ITS primer Methods: The Urban Barcode Project DNA Extraction protocol and procedure. Intro: Truffles are considered a delicacy all around the world and are sold at exorbitant prices. There are numerous kinds of truffles from many different places, but one of the most luxurious truffles is the ‘Perigord’ French Black Truffle (Tuber Melanosporum), also called black diamonds. These truffles are very rare and are only found in a couple places on earth. Another type of truffle is not at all as expensive and is very common. This is the Chinese Black Truffle, which actually consists of three different species: Tuber Sinensis, Tuber Indicum and Tuber Himalayensis. These French and Chinese truffles look very similar, but taste very different and are very differently priced. Chinese Black Truffles can be sold for as little as $20 to $30 a pound, whereas Perigord French Black Truffles start at $800 per pound. As mentioned in a New York Times article by Florence Fabricant, some truffle dealers have been mixing these cheap, and tasteless Chinese Black Truffles with the precious French Black Truffles in order to make a larger profit. The implications and repercussions of this fraud are obvious. To combat this widespread fraud, France’s National Institute of Agronomic Research created a genetic coding test that looks at their DNA to decipher between the two species. Therefore, truffles make an interesting food fraud topic to do DNA barcoding with. Gels/Results: Sampling: The images from the gel electrophoresis did, indeed have bands, but when the DNA was sent to the lab to be sequenced, no DNA match was found. Discussion: When the gels were run, there were both primer dimer and DNA bands indicating that there was viable DNA, but when the DNA was sent to the lab to get sequenced, no results were obtained, which is very strange. PCR conditions are not the same as sequencing conditions, that is probably why some samples amplified nicely and others did not sequence well. One explanation for this trend is that practically all of these samples showed two bands after the gel electrophoresis. It was assumed that one of the bands was a primer dimer band would not interfere with the results of the sample band. However, this assumption may have been misleading, as the primer dimer band may have impeded upon our sample results. When there is a double band two DNA fragments are amplified. These two DNA bands are sequenced at the same time, which causes overlap and creates difficulty in identifying the gene. To avoid this problem in the future one may choose to use different fungal primers in their procedure, increase the annealing temperature in PCR, or add an additional chemical that would inhibit excess 1 Acknowledgements/Bibliography: 1. ● ● ● Melissa Lee’s email explanation ● http: //www. first-nature. com/fungi/tubermelanosporum. php http: //www. eatlovesavor. com/distinguishing-betweenperigord-black-truffles-and-chinese-black-truffles http: //www. nytimes. com/1995/02/15/garden/theinvasion-of-the-chinese-truffle. html http: //www. urbanbarcodeproject. org/files/using-dnabarcodes. pdf